Opa相互作用蛋白5在胰腺癌中的表达及其对PANC-1细胞增殖的影响

目的探索Opa相互作用蛋白5(OIP5)在胰腺癌中的表达及其对PANC-1细胞增殖的影响。方法通过数据库分析OIP5在胰腺癌组织及癌旁组织中的表达;用实时定量PCR(RT-qPCR)和蛋白印迹法(Western blot)分别检测人胰腺癌细胞系MIAPaCa-2、PANC-1、KP-3、BxPC-3细胞中OIP5 mRNA和蛋白表达;构建OIP5基因沉默质粒的慢病毒(pGCSIL-shOIP5)和对照质粒慢病毒(pGCSIL-shCtrl),分别感染PANC-1细胞,分为OIP5基因沉默组和shCtrl对照组,5 d后采用RT-qPCR和Western blot测定慢病毒敲低效率,流式细胞计量术检测细胞凋亡;OIP5基因沉默组和shCtrl对照组连续5 d进行MTT检测和细胞计数;OIP5基因沉默组和shCtrl对照组孵育10 d形成集落,Giemsa染色分别集落总数。结果胰腺癌中OIP5 mRNA表达显著高于正常胰腺组织(P<0.05),OIP5高表达患者的总存活率显著低于OIP5低表达患者(P<0.05),且其无病生存率也显著降低(P<0.05);OIP5在MIAPaCa-2、PANC-1和KP-3中表达较高,而在BxPC-3细胞系中的表达较低;MTT检测结果显示OIP5沉默在第4和第5天显著降低了PANC-1细胞的增殖速率(P<0.01);OIP5沉默后细胞集落数(平均为9个)显著低于shCtrl对照组中的数量(平均为40个)(P<0.01);OIP5沉默后PANC-1细胞凋亡比例为8.3%显著高于shCtrl的4.5%(P<0.01)。结论OIP5在胰腺癌细胞系中异常高表达,OIP5基因可调控胰腺癌PANC-1细胞的增殖、凋亡以及集落形成,提示OIP5可能在胰腺癌发病机制中作为癌基因发挥作用,从而为胰腺癌的靶向治疗提供了潜在的生物标志物。

Expression of Opa interacting protein 5 in pancreatic cancer and its effect on the proliferation of PANC-1 cells

Objective To explore the expression of OPA interacting protein 5(OIP5)in pancreatic cancer and its effect on the proliferation of PANC-1 cells.Methods The expression of OIP5 in pancreatic cancer and adjacent tissues was evaluated by database;the expression of OIP5 mRNA and protein in human pancreatic cancer cell lines like MIAPaCa-2,PANC-1,KP-3 and BxPC-3 was detected by real-time quantitative PCR(RT-qPCR)and Westernblot,respectively;the expression of OIP5 gene silencing plasmid(pGCSIL-shOIP5)and control plasmid(pGCSIL-shCtrl)were observed,respectively infected with PANC-1 cells,then divided into OIP5 gene silencing group and shctrl control group.5 days later,RT-qPCR and Western blot were used to detect slow virus knockdown efficiency,and flow cytometry was used to detect cell apoptosis.MTT and cell count were carried out in OIP5 gene silencing group and shctrl control group for 5 consecutive days.OIP5 gene silencing group and shctrl control group were incubated for 10 days to form colonies,then Giemsa staining was used to count total number of sets.Results The expression of OIP5 mRNA in pancreatic cancer was significantly higher than that in normal pancreatic tissue(P<0.05),and the overall survival rate in patients with high expression of OIP5 was significantly lower than that in patients with low expression of OIP5(P<0.05),and the disease-free survival rate was also significantly lower(P<0.05);the expression of OIP5 was higher in MIAPaCa-2,PANC-1 and KP-3,but lower in BXPC-3 cell line;the results of MTT showed that OIP5 was silenced in the 4 th day.The proliferation rate of PANC-1 cells was significantly decreased on the 4 th and the 5 th day(P<0.01);the number of cell colonies(9 on average)after OIP5 silencing was significantly lower than that in shctrl control group(40 on average)(P<0.01);the apoptosis rate of PANC-1 cells after OIP5 silencing was 8.3%and 4.5%higher than that of shctrl(P<0.01).Conclusions OIP5 is highly expressed in pancreatic cancer cell lines.OIP5 gene can regulate the proliferation,apoptosis and clonal formation of PANC-1 cells,suggesting that OIP5 may play a role as a oncogene in the pathogenesis of pancreatic cancer,thus providing a potential biomarker for targeted treatment of pancreatic cancer.